FastDigest NheI

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	The amplified PCR product was digested with SacI at the 5′ and with NheI at the 3′ end followed by ligated into SacI+NheI (FastDigest®, Fermentas, USA) digested pSY-B-MCS plasmid to yield a construct pSY-B-GFP possessing GFP gene between the MP and CP region (Fig. 2A).	Recombinant clones were confirmed through colony PCR with GFP gene-specific primers, restriction digestion and nucleotide sequencing.

Protocol tips
The amplified PCR product was digested with SacI at the 5′ and with NheI at the 3′ end followed by ligated into SacI+NheI (FastDigest®, Fermentas, USA) digested pSY-B-MCS plasmid to yield a construct pSY-B-GFP possessing GFP gene between the MP and CP region (Fig. 2A).
Downstream tips
Recombinant clones were confirmed through colony PCR with GFP gene-specific primers, restriction digestion and nucleotide sequencing.

Publication protocol

"The coding region of green fluorescent protein (GFP,
756 bp) was amplified by PCR from the plasmid pCAMBIA 1302 binary vector (Accession No.: AF234298) using
a primer pair RKJ268F and RKJ277R and Phusion® HighFidelity DNA polymerase (Table 1). The amplified PCR
product was digested with SacI at the 5′ and with NheI at
the 3′ end followed by ligated into SacI+NheI (FastDigest®,
Fermentas, USA) digested pSY-B-MCS plasmid to yield a
construct pSY-B-GFP possessing GFP gene between the MP
and CP region (Fig. 2A). Recombinant clones were confirmed through colony PCR with GFP gene-specific primers,
restriction digestion and nucleotide sequencing"

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